U.S. patent application number 12/412598 was filed with the patent office on 2009-10-01 for therapeutic composition for atopic dermatitis.
Invention is credited to Masahiko KURODA, Katsuko Sudo, Masakatsu Takanashi, Shigeki Yamauchi.
Application Number | 20090246181 12/412598 |
Document ID | / |
Family ID | 40565058 |
Filed Date | 2009-10-01 |
United States Patent
Application |
20090246181 |
Kind Code |
A1 |
KURODA; Masahiko ; et
al. |
October 1, 2009 |
THERAPEUTIC COMPOSITION FOR ATOPIC DERMATITIS
Abstract
A novel type of therapeutic agent, human mesenchymal stem cells,
and a composition containing the same for the treatment of atopic
dermatitis is disclosed. A method for the treatment of atopic
dermatitis in patient is also disclosed.
Inventors: |
KURODA; Masahiko; (Tokyo,
JP) ; Takanashi; Masakatsu; (Kanagawa, JP) ;
Sudo; Katsuko; (Saitama, JP) ; Yamauchi; Shigeki;
(Kanagawa, JP) |
Correspondence
Address: |
MILLEN, WHITE, ZELANO & BRANIGAN, P.C.
2200 CLARENDON BLVD., SUITE 1400
ARLINGTON
VA
22201
US
|
Family ID: |
40565058 |
Appl. No.: |
12/412598 |
Filed: |
March 27, 2009 |
Current U.S.
Class: |
424/93.7 ;
435/366; 435/372 |
Current CPC
Class: |
A61K 2035/124 20130101;
A61P 37/08 20180101; A61P 17/00 20180101; C12N 5/0663 20130101 |
Class at
Publication: |
424/93.7 ;
435/366; 435/372 |
International
Class: |
A61K 35/28 20060101
A61K035/28; C12N 5/08 20060101 C12N005/08; A61P 17/00 20060101
A61P017/00 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 28, 2008 |
JP |
2008-088641 |
Claims
1. A composition for the treatment of atopic dermatitis comprising
human mesenchymal stem cells as the active principle.
2. The composition for the treatment of atopic dermatitis according
to claim 1, wherein the atopic dermatitis is that which has
developed g in a mammal.
3. The composition for the treatment of atopic dermatitis according
to claim 2, wherein the mammal is selected from the group
consisting human, dog, cat, rabbit, and rodent.
4. The composition for the treatment of atopic dermatitis according
to claim 1, wherein the human mesenchymal stem cells are of
human-bone marrow origin.
5. The composition for the treatment of atopic dermatitis according
to claim 4, wherein the mammal is a human and is not the same as
the human from whom the human mesenchymal stem cells originate.
6. The composition for the treatment of atopic dermatitis according
to claim 1, wherein the composition is an injection.
7. The composition for the treatment of atopic dermatitis according
to claim 6, wherein the composition is an injection for intravenous
administration.
8. A method for the treatment of atopic dermatitis in a patient
comprising administering to the patient a therapeutically effective
amount of human mesenchymal stem cells.
9. The method for the treatment of atopic dermatitis in a patient
according to claim 8, wherein the patient is a mammal.
10. The method for the treatment of atopic dermatitis in a patient
according to claim 9, wherein the mammal is selected from the group
consisting of human, dog, cat, rabbit, and rodent.
11. The method for the treatment of atopic dermatitis in a patient
according to claim 8, wherein the human mesenchymal stem cells are
of human bone-marrow origin.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to a therapeutic composition
for atopic dermatitis, more specifically to a therapeutic
composition for dermatitis comprising, as an active principle,
mesenchymal stem cells of a mammal, in particular of a human.
BACKGROUND OF THE INVENTION
[0002] IgE, an isotype of immunoglobulin, is produced by such B
cells that have been stimulated with an antigen and then switched
into IgE class cells, as being specific to the very antigen. IgE
molecules thus produced then bind at their Fc region to Fc.epsilon.
receptors present on the surface of mast cells, which occur in
subcutaneous as well as connective tissues, or of basophils, and
thus stay there. If they come into contact with the same antigen,
two of adjacent IgE molecules bind to the antigen falling between
them, via which two adjacent Fc.epsilon. receptors thus become
cross-linked. This triggers activation of certain membrane-bound
enzymes. This activation results, through degranulation, in an
abrupt release of chemical mediators such as histamine that are
stored in numerous granules within those cells, as well as in the
release of other chemical mediators from the cell membrane, such as
leukotrienes, prostaglandins, platelet activating factors, and the
like, thereby bringing about a wide range of physiological
effects.
[0003] Atopy is a diathesis with which production of IgE is readily
induced even against those common antigens which generally occur in
the environment and induce no particular immune response in healthy
individuals. The body of an individual with this diathesis, whether
a human or other mammals, is predisposed to recognize as allergens
some or other of such factors in the environment that are generally
harmless, and thus produces IgE which is specific to them. The IgE
thus produced is accumulated on the surface of mast cells and like
cells. When it is contacted by the same antigen, release from the
cells of histamine and other chemical mediators is triggered.
Histamine has such activities as a bronchial smooth muscle
contracting activity, a blood vessel permeability enhancing
activity, and a mucus secretion inducing activity, which, in
combination with the activities of other chemical mediators, bring
about a series of allergic reactions, causing damages to various
tissue, too (atopic dermatitis, atopic type bronchial asthma,
allergic rhinitis, and the like).
[0004] Atopic dermatitis, one of atopic diseases, formerly
developed in early childhood in most patients, and cured before
they grew into adults. In recent year, however, the number of the
cases has been increasing in which cure is not attained even though
the patient has grown into an adult, and in increasing number of
patients, the disease which once cured, recurs in his or her
adulthood, or the disease first develops only after the patient has
reached his or her adulthood. As a result, the total number of
patients with atopic dermatitis has been rapidly increasing; from
318,000 in 1996, to 399,000 in 1999, and at present greatly
expanding toward 500,000 (estimated by Ministry of Health, Labor
and Welfare).
[0005] Atopic dermatitis first develops on the head of the patient
in majority of the cases, and then gradually expands to the face,
the earlobes, the trunk, and the limbs. Its symptom is chronic
eruptions in the skin (rashes), and in accordance with the degree
of its progression and the age of the patient, exhibits a wide
variety of conditions including, for example, erythema
(swelling/edema/ infiltration, lichenification), papules (solid,
serous), pruriginous nodules, scales (pityroid, foliate,
figuratus), crusts (scabs), blisters, pustula, erosion, ulcer,
pigmentation, depigmentation, xerosis, and pore cornification,
etc., with persistent severe itching. As the disease is aggravated
into severe stages, such symptoms are noted as erythema accompanied
by severe swelling/edema/infiltration or lichenification, increased
formation of papules, severe grades of scales, adhered crusts,
small blisters, erosion, a number of scratch wound traces and
pruriginous nodules, and thereby the quality of life (QOL) of the
patient is heavily impaired.
[0006] As atopic dermatitis exhibits severe symptoms like these,
the increase in the number of its patient is becoming a social
issue. Since a drug therapy has not been found yet that could bring
about complete cure of the disease, there is no other choice at
present than to resort to symptomatic therapies. The drugs whose
efficacy and safety have so far been established for easing
inflammation in atopic dermatitis are external steroidal
preparations. They are classified according to their potency of
effects: for severely affected patients, the first choice is
considered to be those external preparations which exhibit most
potent effects and thus belong to the so-called "strongest" class
(such as clobetasol propionate, diflorasone diacetate), or those
which follow next to them and belong to the "very strong" class
(such as mometasone furoate, betamethasone butyrate propionate,
fluocinonide, betamethasone dipropionate, difluprednate,
budesonide, amcinonide, diflucortolone valerate, hydrocortisone
butyrate propionate). For patients who are moderately affected,
external steroidal preparations of the "strong" class, or those of
the "medium" class with moderate potency of effects, are considered
to be the first choice, and for patients who are affected only
mildly, external steroidal preparations of the "medium" class or
less potent ones.
[0007] These external steroidal preparations, however, are nothing
more than drugs for symptomatic treatment, and therefore they must
be used continuously to obtain persistent suppression of atopic
dermatitis. And in these drugs, the potency of their activity and
that of their side effects generally correlate with each other. For
example, if an external steroidal preparation of the "strongest"
class is applied at high doses and for an extended length of time,
side effects will sometimes result due to the systemic absorption
of the drug, such as suppression of the functions of the
pituitary-adrenal cortex system or induction of the development of
cataract or glaucoma. Furthermore, suppression of cell growth by
steroids could cause such disorders in the skin area to which they
are applied for an extended length of time, as dermatrophy, striae
cutis distensae, dryness, ichthyosis-like lesion, retardation of
wound healing, steroid purpura, telangiectasis, dyschromatosis,
rosacea-like dermatitis, etc. For theses reasons, it sometimes is
difficult to continuously apply external steroidal preparations for
an extended period of time. It also is a problem that rebound would
occur when application of them is disrupted after an extended
period of use. Furthermore, not a small number of people mistrust
steroids, and they, avoiding external steroidal preparations, have
failed to receive even a necessary and proper treatment until their
conditions become severe, thereby suffering a great
disadvantage.
[0008] Thus, while the number of the patients has been great and
even increasing further, there has so far been no such therapeutic
composition for atopic dermatitis that is sufficient in both
aspects of efficacy and safety. Thus, development of a new
therapeutic composition for the treatment of atopic dermatitis has
been longed for.
[0009] Mesenchymal stem cells (MSC) are multipotent stem cells
which occur in a very small number in bone marrow and other
mesenchymal tissues. They have high potential to proliferate and
are capable of differentiating into osteocytes, chondrocytes,
muscle cells, tendon cells, stromal cells, adipose cells, and the
like. It is known that human mesenchymal stem cells can be cultured
in an artificial medium to let them proliferate (see Patent
Document 1), and, like other cultured cells, can be stored and
supplied in a frozen state. It is known, too, that mesenchymal stem
cells can be obtained not only from the bone marrow but also from
various other tissues such as adipose tissue (see Patent Document
2), dental pulp tissue (see Patent Document 3), placenta tissue,
umbilical cord tissue (see Patent Document 4), and the like.
[0010] Disclosures have been made that rejection of an allogeneic
skin graft between baboons is suppressed in the recipient which
have been administered with baboon mesenchymal stem cells; that
graft-versus-host disease (GvHD) in a dog after allogeneic
transplantation of bone marrow is suppressed by administration of
canine mesenchymal stem cells; that human T lymphocytes do not show
proliferative response to human (allogeneic) mesenchymal stem cells
in vitro (mixed lymphocyte reaction); that human mixed lymphocyte
reaction is suppressed non-specifically to MHC type by human
mesenchymal stem cells, and the same is observed in dogs; and, on
the basis of these results, about the possibility of applying human
mesenchymal stem cells for suppression of immune responses after
transplantation between humans (see Patent Documents 5, 6, which
derive from a common priority application, and Patent Document 7,
which is a translation of the latter).
[0011] Expecting such effects of mesenchymal stem cells as
mentioned above, a clinical trial was conducted in the United
States on steroid-resistant severe GvHD patients to evaluate human
mesenchymal stem cell's suppressive effect on acute
graft-versus-host disease (GvHD) in human following bone marrow
transplantation, and the result has been reported (see Non-patent
Documents 1 and 2). According to the report, the patients had been
treated in advance with anticancer compositions (cyclophosphamide,
busulphan, fludarabin) and/or with total body irradiation (TBI)
against their malignant tumors, and were administered with
methotrexate, cyclophosphamide, or prednisolone for prevention and
treatment of GvHD, and, under those conditions, the patients were
administered with human mesenchymal stem cells
(0.7-9.times.10.sup.6 cells/kg body weight). The administration of
the human mesenchymal stem cells to the patients, who had their
immune system suppressed, was reported to have induced a response
in six out of the eight patients as a whole.
[0012] In connection with GvHD, though mesenchymal stem cells are
thought to bring about T cells-mediated suppression of the immune
cells of the host (see Non-patent Document 3), the clear mechanism
of it is yet to be elucidated.
[0013] It is also proposed that mesenchymal stem cells be used to
treat neovascularization, autoimmune diseases, inflammatory
responses (in Alzheimer disease, Parkinson's disease, cerebral
stroke, brain-cell lesions, psoriasis, chronic dermatitis, contact
dermatitis, arthrosteitis, arthritis including rheumatoid
arthritis, inflammatory bowel disease, chronic hepatitis), cancer,
allergic diseases, sepsis, trauma (burn, surgery, transplantation),
inflammation of various tissues or organs (cornea, lens, pigment
epithelium, retina, brain, spinal cord, uterus during pregnancy,
ovary, testis, adrenal gland) (see Patent Document 8). However, in
this document, the functions of mesenchymal stem cells were
examined in vitro only, and no examination was conducted either in
vivo or in a model of any of those particular diseases. Therefore,
the document does not provide any result which could serve as a key
to evaluating usefulness of mesenchymal stem cells for such
diseases. [0014] [Patent Document 1] U.S. Pat. No. 5,486,359 [0015]
[Patent Document 2] Japanese Patent Application Publication
2004-129549 [0016] [Patent Document 3] Japanese Patent Application
Publication 2004-201612 [0017] [Patent Document 4] Japanese Patent
Application Publication 2004-210713 [0018] [Patent Document 5] U.S.
Pat. No.6,328,960 [0019] [Patent Document 6] WO 99/47163 [0020]
[Patent Document 7] Japanese Patent Application Publication
2002-50683 [0021] [Patent Document 8] WO 2005/093044 [0022]
[Non-patent Document 1] Transplantation. 2006; 81(10): 1390-7
[0023] [Non-patent Document 2] Br J Haematol. 2007; 137(2): 87-98
[0024] [Non-patent Document 3] Blood. 2005; 105(4): 1815-22
SUMMARY OF THE INVENTION
[0025] Under the above circumstances, the objective of the present
invention is to provide a non-steroidal, novel type of composition
for the treatment of atopic dermatitis.
BRIEF DESCRIPTION OF THE FIGURES
[0026] The present inventors injected atopic dermatitis model mice
intravenously with human mesenchymal stem cells, and followed their
course of disease. It was found that the mice did not exhibit any
abnormal reactions even though the injected cells were heterologous
to them, and that not only the progress of their dermatitis was
greatly suppressed after just a limited times of administration,
but also the condition of their skin rapidly improved toward
normalization. The present invention was completed based upon those
findings.
[0027] Thus, the present invention provides what follows:
[0028] 1. A composition for the treatment of atopic dermatitis
comprising human mesenchymal stem cells as the active
principle.
[0029] 2. The composition for the treatment of atopic dermatitis
according to 1 above, wherein the atopic dermatitis is that which
has developed in a mammal.
[0030] 3. The composition for the treatment of atopic dermatitis
according to 2 above, wherein the mammal is selected from the group
consisting human, dog, cat, rabbit, and rodent.
[0031] 4. The composition for the treatment of atopic dermatitis
according to one of 1 to 3 above, wherein the human mesenchymal
stem cells are of human bone-marrow origin.
[0032] 5. The composition for the treatment of atopic dermatitis
according to 4 above, wherein the mammal is a human and is not the
same as the human from whom the human mesenchymal stem cells
originate.
[0033] 6. The composition for the treatment of atopic dermatitis
according to one of 1 to 5 above, wherein the composition is an
injection.
[0034] 7. The composition for the treatment of atopic dermatitis
according to 6 above, wherein the composition is an injection for
intravenous administration.
[0035] 8. A human mesenchymal stem cell for use as a medicament for
the treatment of atopic dermatitis.
[0036] 9. The human mesenchymal stem cell according to 8 above,
wherein the atopic dermatitis is mammal's atopic dermatitis.
[0037] 10. The human mesenchymal stem cell according to 9 above,
wherein the mammal is selected from the group consisting of human,
dog, cat, rabbit, and rodent.
[0038] 11. The human mesenchymal stem cell according one of 8 to 10
above, wherein the human mesenchymal stem cell is of human
bone-marrow origin.
[0039] 12. The human mesenchymal stem cell according to one of 8 to
11 above, wherein the mammal is a human.
[0040] 13. The human mesenchymal stem cell according to 12 above,
wherein the human is not the same as the human from whom the
mesenchymal stem cell originates.
[0041] 14. A method for the treatment of atopic dermatitis in a
patient comprising administering to the patient a therapeutically
effective amount of human mesenchymal stem cells.
[0042] 15. The method for the treatment of atopic dermatitis in a
patient according to 14 above, wherein the patient is a mammal.
[0043] 16. The method for the treatment of atopic dermatitis in a
patient according to 14 or 15 above, wherein the mammal is selected
from the group consisting of human, dog, cat, rabbit, and
rodent.
[0044] 17. The method for the treatment of atopic dermatitis in a
patient according to one of 14 to 16 above, wherein the human
mesenchymal stem cells are of human bone-marrow origin.
[0045] Human mesenchymal stem cells, and therefore the composition
for the treatment of atopic dermatitis according to the present
invention, can not only remarkably suppress the progression of
atopic dermatitis in a mammal, but also greatly improve the
condition of the skin. Further, they exhibit such effects after
only a very limited number of rounds of their administration, and
persistently thereafter. Human mesenchymal stem cells and the
composition for the treatment of atopic dermatitis according to the
present invention, therefore, can be used with great advantages for
the treatment of atopic dermatitis in a human or a pet (dog, cat,
rodent, rabbit, or the like).
[0046] Thus, present invention also provides use of human
mesenchymal stem cells for the production of a composition for the
treatment of atopic dermatitis in a patient, as well as a method
for the treatment of atopic dermatitis in a patient, e.g., mammal,
such as human, dog, cat, rabbit, rodent, and the like.
BRIEF DESCRIPTION OF THE FIGURES
[0047] FIG. 1 is a photograph showing the general appearance of an
atopic dermatitis model mouse that has developed score 4
dermatitis.
[0048] FIG. 2 is a photograph showing the general appearance of a
mouse before developing dermatitis.
[0049] FIG. 3 is a graph showing the profiles of the dermatitis
scores in the MSC-administered and the control groups.
DETAILED DESCRIPTION OF THE INVENTION
[0050] The composition for the treatment of atopic dermatitis
according to the present invention comprises human mesenchymal stem
cells as the active principle (active agent). In the present
invention, any human mesenchymal stem cells may be employed that
are taken from any tissue including bone marrow, adipose tissue,
dental pulp tissue, placenta tissue, etc. Preferable, however, are
those originating from human bone marrow. Mesenchymal stem cells
which were obtained from human bone marrow and formed into a cell
line are already on the market (Poetics.TM., Cambrex Bio Science
Walkersville, Inc., MD, USA), and may be employed, either directly
or after further subcultivation.
[0051] In the present invention, it is not necessary for the human
mesenchymal stem cells to be of a primary culture, but subcultured
cells may be employed. While primary culture cells may be used
directly, those usually used may be subcultured cells, considering
the actual requirements for convenience in their production through
cell growth, as well as their storage and use, in a substantial
amount. In the present invention, the active principle, human
mesenchymal stem cells, may be supplied, for example, in a frozen
state, and thawed just before their administration, suspended in an
aqueous medium, and then administered. However, doing so is not a
prerequisite and, for example, human mesenchymal stem cells
collected from a culture and not having undergone freezing, may be
used directly in the form of a suspension. In doing this, an
aqueous medium in which to suspend the cells may be, for example,
an injectable aqueous solution which has its osmotic pressure, pH,
and salts concentration, and the like adjusted substantially to
those of the blood. Intravenous fluids such as acetated Ringer's
solution, glucose acetated Ringer's solution, etc., physiological
saline, or glucose solution are non-limiting examples which may be
used. For example, to Ringer's solution for intravenous infusion
which is to be used, an acceptable amount of dimethylsulfoxide
(DMSO) or human serum albumin (HSA) may be added.
[0052] In the present invention, human mesenchymal stem cells may
be administered in the form of an injectable preparation, whether
intravenously, intramuscularly, or subcutaneously, etc., among
which intravenous administration is preferred. When conducting
intravenous administration, the injectable preparation may be
administered either directly from the syringe to the patient, or
dropwise from a drip bag containing the intravenous fluid to which
its has been added beforehand. Furthermore, human mesenchymal stem
cells may be injected directly in the affected area of the
skin.
[0053] As to the administration regimen of the composition
according to the present invention, administration may be made only
once, or it may be made multiple times, e.g., twice, thrice or
more, until the effect of composition is observed, according to the
severity of the symptoms of atopic dermatitis. However, since the
composition according to the present invention exhibits remarkable
persistent effect after administration, and, further, as the effect
of the composition according to the present invention is thought to
consist in inducing autonomous normalization of the behavior of the
patient's own immune system in the skin, no additional
administration of the composition will be needed as long as the
inflammation has been ameliorated and the condition is
continuing.
[0054] The composition for the treatment of atopic dermatitis
according to the present invention is a therapeutic composition
commonly applicable to any patient, and thus it will be
administered, in general, to patients who are allogeneic to the
origin of the human mesenchymal stem cells contained as the active
principle. However, its administration to an syngeneic patient
(e.g., one of the homozygotic twins the other of whom is the origin
of the mesenchymal stem cells) is also permitted. Further, it is
also allowed that the composition for the treatment of atopic
dermatitis according to the present invention, which has, through
the process of cell growth in culture, been prepared as a commonly
applicable composition, is administered by chance to the very
patient who is the origin of the mesenchymal stem cells.
[0055] When administered, the density of the human mesenchymal stem
cells in the composition according to the present invention may be
preferably 1.times.10.sup.2-1.times.10.sup.9 cells/mL, and more
preferably 1.times.10.sup.3-1.times.10.sup.8 cells/mL. And the
number of the cells to be administered to a human at one time may,
in general, be 1.times.10.sup.5-1.times.10.sup.7 cells/kg body
weight, though it depends on the intended frequency of
administration. However, these are not a prerequisite, and those
numbers may altered as desired in accordance with the degree of the
symptoms. Particularly, according to the findings that the
inventors have already obtained separately through administration
of mesenchymal stem cells to arthritis model animals (Japanese
Patent Application No. 2007-233094, not yet laid open), human
mesenchymal stem cells administered to an animal homed to the site
of inflammation, and, therefore, even if the number of human
mesenchymal stem cells administered is relatively small, they
accumulate at the site of inflammation and exhibit an excellent
effect there. Also, human mesenchymal stem cells, when they are no
longer required, will vanish away by themselves. Thus, the dose of
human mesenchymal stem cells may be set as desired in a wide
range.
EXAMPLE 1
[0056] The present invention will be described in further detail
below with reference to examples. However, it is not intended that
the present invention be limited to those examples. The method for
making atopic dermatitis model mice, the schedule of administration
of mesenchymal stem cells, and the method of evaluation of
dermatitis are presented below.
[Method for Making Atopic Dermatitis Model Mice]
[0057] NC/Nga mice are mice of a pure strain that have been derived
from an indigenous Japanese mice called "Nishiki Nezumi". NC/Nga
mice will not develop dermatitis if maintained under an SPF
(Specific Pathogen Free) condition. However, if they are maintained
under a conventional, i.e., a non-SPF condition, they will develop
dermatitis. The dermatitis thus developed appears, both clinically
and histologically, very similar to human atopic dermatitis,.
Namely, the mice exhibit the symptom of strong itchiness, rougher
body hair, especially on the head and the back, erythema,
eczema-like lesions, hemorrhage, crusts, excoriation, erosions,
scaling and dryness, which occur everywhere in the body surface
including the ears, nose, and dorsal skin. NC/Nga mice maintained
under a non-SPF condition have higher blood IgE values than those
maintained under a SPF condition, and exhibit infiltration of
numerous eosinophils in the skin, increase in number of mast cells,
and degranulation by their activation. The symptoms will progress
and become severe along with the weeks of age, which sometimes goes
to the loss of the earlobe (Int. Arch. Allergy Immunol., 120
(Supple. 1), 70-75). Dermatitis similar to atopic dermatitis is
also inducible in these mice, by letting mites parasitize them, or
also by application of a drug such as picryl chloride (Biol. Pharm.
Bull. 28(1) 78-82 (2005)), and thus they are used as atopic
dermatitis model mice.
[0058] Male, four-week old NC/Nga mice (Japan Slc, Inc., Hamamatsu,
Shizuoka) were purchased and maintained for two weeks under a SPF
condition for acclimatization before they were used in the test.
Throughout this period of acclimatization and testing, they were
maintained under the condition where lighting was on and off at
12-hour intervals, and the temperature was kept constant, with
.gamma.-irradiated food and water provided, to which the mice was
accessible ad libitum. The method for keeping them complied with a
relating rule of Experimental Animals Center, Tokyo Medical
University. After the acclimatization, the mice were parasitized by
Myobia musculi, and maintained for further 10-12 weeks to develop
spontaneous dermatitis. The general condition of the mice were
observed, and the severity of their dermatitis was evaluated
according to the criterion for evaluation of severity of dermatitis
as shown in Table 1. Six mice whose dermatitis symptoms were graded
as score 2 (moderate) were taken out and divided into two groups,
thee mice each.
TABLE-US-00001 TABLE 1 Criterion for Evaluation of Severity of
Dermatitis Score Symptoms 0 (normal) no dermatitis, or complete
cure 1 (mild) very mild erythema, mild wound, or cure to comparable
grade 2 (moderate) manifest erythema, wound, mild hemorrhage, or
cure to comparable grade 3 (severe) sever erythema, wound (partial
loss of earlobe), localized ulcer, crust formation, or cure to
comparable grade 4 (loss of earlobe) much hemorrhage, or loss of
earlobe with hemorrhage, wide range of ulcer
[Administration of Human Mesenchymal Stem Cells]
[0059] In one (MSC administered group, n=3) of the two groups of
atopic dermatitis model mice prepared above, the mice were
administered with 1.times.10.sup.6 cells of human mesenchymal stem
cells (Poietics.TM., Cambrex: bone marrow-derived, CD105+,
CD166+,CD29+, CD44+, CD14-, CD34-, CD45-), which had been suspended
in 0.21 mL of acetated Ringer's solution for intravenous infusion
fluid (PlasmaLyteA:Baxter) containing 3.7% DMSO and 5% HSA (human
serum albumin) and been kept at room temperature, through tail vein
at 1 mL/min. After the mice was warmed over a warm bath to let the
vein dilate, the administration was carried out to constrained mice
put in a fixation device. Following the first administration of
human mesenchymal stem cells (day 1), administration of human
mesenchymal stem cells was repeated in the same manner also on days
2 and 3. In the other group (Control group, n=3), the mice
received, in the same manner as above, 0.21 mL of acetated Ringer's
solution for intravenous infusion fluid containing 3.7% DMSO and 5%
HSA, which was free of human mesenchymal stem cells.
[Evaluation of Dermatitis]
[0060] Severity of dermatitis was evaluated according to the
criterion presented in Table 1. Evaluation was made everyday, from
day 1 through day 10. The dermatitis score for each animal and the
mean score for each group is given in Table 2.
TABLE-US-00002 TABLE 2 Dermatitis Score Control group MSC
administered group No. 1 No. 2 No. 3 Mean No. 1 No. 2 No. 3 Mean
Day 1 2 2 2 2 Day 1 2 2 2 2 Day 2 3 3 3 3 Day 2 3 3 3 3 Day 3 4 3 4
4 Day 3 3 3 3 3 Day 4 4 4 4 4 Day 4 3 3 3 3 Day 5 4 4 4 4 Day 5 3 3
3 3 Day 6 4 4 4 4 Day 6 3 3 3 3 Day 7 4 4 4 4 Day 7 3 3 2 3 Day 8 4
4 4 4 Day 8 2 2 2 2 Day 9 4 4 4 4 Day 9 2 2 1 2 Day 10 4 4 4 4 Day
10 1 2 1 1
[0061] In both groups, the dermatitis score was 2 for all the
animals on day 1. In the control group, however, on day 2 (at the
second administration), the score was 3 for all the animals; on day
3, one of the three animals had score 3 but the other two had the
highest score of 4; and on day 4, all the animals had score 4,
resulting in severe dermatitis. As a result, in the control group,
there were observed much hemorrhage from the earlobe or partial
loss of the earlobe with hemorrhage, and ulcer ranging from the
head, face, earlobe to trunk and forelimbs. All the animals in the
control group subsequently constantly exhibited the severest, score
4 dermatitis. FIG. 1 shows the general appearance of a mouse before
the development of dermatitis, and FIGS. 2 the general appearance
of mouse No. 1 in the control group, on day 3, respectively.
[0062] In contrast, though the severity of dermatitis in the MSC
administered group was graded as score 2 for all the animals on day
1, and then score 3 for all the animals on day 2, no aggravation to
score 4 dermatitis occurred thereafter in any of the animals, and,
on the contrary, since 4 days after the last administration (day 3)
of MSC, start of remarkable improvement of the skin condition was
observed. Namely, the severity of dermatitis in all the animals of
the MSC administered group stayed at score 3 from day 4 through day
6; the score then was improved to 2 in one of the three animals on
day 7; score 2 in all the three animals on day 8; score 1, which
was lower than the score recorded when MSC administration was
started, in one of the three animals on day 9; and score 1 in also
one of the remaining two animals on day 10. FIG. 3 shows a graphic
illustration of the profile mean value of the scores for each
group.
[0063] As evident from the results, in MSC-administered animals,
not only formation of severe dermatitis was prevented, but also
rapid improvement of the skin condition followed, indicating that
administration with human mesenchymal stem cells exhibited
remarkable therapeutic effect on atopic dermatitis. Especially, the
skin condition remarkably improved after the administration of MSC
on day 1 through day 3 was over, i.e., 4 days after the termination
of MSC administration (thus, from day 7 on), which strongly
suggests that the administered MSC acted, by some mechanism, to
autonomously normalize the behavior of the immune system in the
skin. Meanwhile, nothing abnormal was observed in general
appearance, such as an adverse immune reaction, in any of the mice
of the MSC group, which received repeated (thee times)
administration of the heterologous, human MSC.
Preparation Example 1 Injection
TABLE-US-00003 [0064] Human mesenchymal stem cells (of bone-marrow
origin) 5 .times. 10.sup.7 cells Acetated Ringer's solution to 5
mL
[0065] Human mesenchymal stem cells are suspended in acetated
Ringer's solution, and the total volume of the suspension is
adjusted to 5 mL to make an injection.
Preparation Example 2 Injection
TABLE-US-00004 [0066] Human mesenchymal stem cells (of bone-marrow
origin) 1 .times. 10.sup.8 cells Sterilized physiological saline to
10 mL
[0067] Human mesenchymal stem cells are suspended in sterilized
physiological saline, and the total volume of the suspension is
adjusted to 10 mL to make an injection.
Preparation Example 3 Injection
TABLE-US-00005 [0068] Human mesenchymal stem cells (of bone-marrow
origin) 2 .times. 10.sup.6 cells 5% glucose solution to 2 mL
[0069] Human mesenchymal stem cells are suspended in 5% glucose
solution, and the total volume of the suspension is adjusted to 2
mL to make an injection.
INDUSTRIAL APPLICABILITY
[0070] The composition for the treatment of atopic dermatitis
according to the present invention, which contains human
mesenchymal stem cells as the active agent, can not only suppress
the progression of atopic dermatitis in a mammal but also can
remarkably improve the condition of the skin, and, further,
persistently achieves the effect after only a very limited number
of rounds of administration. Therefore, the active agent is highly
useful as a novel type of therapeutic agent for the treatment of
atopic dermatitis in a human or a mammalian pet.
[0071] Without further elaboration, it is believed that one skilled
in the art can, using the preceding description, utilize the
present invention to its fullest extent. The preceding preferred
specific embodiments are, therefore, to be construed as merely
illustrative, and not limitative of the remainder of the disclosure
in any way whatsoever.
[0072] The entire disclosures of all applications, patents and
publications, cited herein and of corresponding Japanese
application No. 2008-088641, filed Mar. 28, 2008, are incorporated
by reference herein.
[0073] The preceding examples can be repeated with similar success
by substituting the generically or specifically described reactants
and/or operating conditions of this invention for those used in the
preceding examples.
[0074] From the foregoing description, one skilled in the art can
easily ascertain the essential characteristics of this invention
and, without departing from the spirit and scope thereof, can make
various changes and modifications of the invention to adapt it to
various usages and conditions.
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